Beta-Defensin 124 Is Required for Efficient Innate Immune Responses in Prostate Epithelial RWPE-1 Cells.
10.4111/kju.2014.55.6.417
- Author:
Kyeoung Hwa KIM
1
;
Jaehyouk LEE
;
Jun Hyun HAN
;
Soon Chul MYUNG
Author Information
1. Research Institute for Translational System Biomics, Chung-Ang University College of Medicine, Seoul, Korea. uromyung@cau.ac.kr
- Publication Type:Original Article
- Keywords:
Chemotaxis;
Cytokines/chemokines;
Defensins;
Innate immunity;
NF-kappa B
- MeSH:
Bacterial Infections;
Blotting, Western;
Chemokines;
Chemotaxis;
Chromatin Immunoprecipitation;
Cytokines;
Defensins;
Enzyme-Linked Immunosorbent Assay;
Humans;
Immunity, Innate*;
Immunohistochemistry;
Monocytes;
NF-kappa B;
Peptidoglycan;
Phosphorylation;
Prostate*;
Real-Time Polymerase Chain Reaction;
Up-Regulation
- From:Korean Journal of Urology
2014;55(6):417-425
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: The present study aimed to determine the role played by beta-defensin 124 (DEFB124) in the innate immunity of prostate epithelial RWPE-1 cells during bacterial infection. MATERIALS AND METHODS: The expression of DEFB124 was examined by quantitative real-time polymerase chain reaction (PCR), Western blotting, and immunocytochemistry. Enzyme-linked immunosorbent assays and quantitative real-time PCR were performed to determine the production of cytokines and chemokines. Western blotting and chromatin immunoprecipitation studies were performed to assess the interaction between DEFB124 and nuclear factor-kappa B (NF-kappaB) in peptidoglycan (PGN)-stimulated RWPE-1 cells. By chemotaxis assay, we assessed the effect of DEFB124 on the migration of monocytes. RESULTS: Exposure to PGN induced DEFB124 upregulation and NF-kappaB activation through IkappaBalpha phosphorylation and IkappaBalpha degradation. Bay11-7082, an NF-kappaB inhibitor, blocked PGN-induced DEFB124 production. Also, NF-kappaB was shown to be a direct regulator and to directly bind to the -3.14 kb site of the DEFB124 promoter in PGN-treated human prostate epithelial RWPE-1 cells. When DEFB124 was overexpressed in RWPE-1 cells, interestingly, the production of cytokines (interleukin [IL] 6 and IL-12) and chemokines (CCL5, CCL22, and CXCL8) was significantly increased. These DEFB124-upregulated RWPE-1 cells markedly induced chemotactic activity for THP-1 monocytes. CONCLUSIONS: Taken together, these results provide strong evidence for the first time that increased DEFB124 expression via NF-kappaB activation in PGN-exposed RWPE-1 cells enhances the production of cytokines and chemokines, which may contribute to an efficient innate immune defense.
